For parthenogenetic activation as a model system of nuclear transfer, microinjection and electroporation as activation treatments in bovine metaphase II oocytes were administered to each of three groups as follows: control group (treatments with Ca2+, Mg2+ -free PBS+100 micro M EGTA), IP3 group (control+25 micro M IP3) and IP3+ ryanodine group (control+25 micro M IP3+10 mM ryanodine). In experiments using microinjection, no significant differences were observed between any of the developmental stages of the electroporation experiment. For electroporation, cleavage rates were significantly higher in the IP3+ryanodine group than in the IP3 or control group (85.6% vs 73.7% or 67.6%, respectively). In the subsequent stages of embryonic development, such as morula and blastocyst formation, the IP3 and ryanodine group exhibited significantly higher rates of morula fomation than the IP3 or control groups (40.6% vs 24.2% or 16.7%, respectively). Similarly, the rate of blastocyst formation in the IP3+ryanodine group was significantly higher than the control group (16.3% vs 6.9%) but did not differ significantly from the IP3 group (16.3% vs 9.5%). In nuclear transfer, activation was performed at 30 hpm by microinjection and elecroporation with 25 micro M IP3+ 10 mM ryanodine followed by 6-DMAP treatment. No significant differences were observed at any stage of embryonic development and none of the embryos activated by electroporation reached either the morula or blastocyst stage. However, 3.8% and 1.9% of embryos activated by microinjection sucessfully developed to the morula and blastocyst stages, respectively. In conclusion, activation treatments using IP3 and ryanodine are able to support the development of bovine parthenogenetic and reconstructed embryos.
Adenine/administration & dosage/*analogs & derivatives/pharmacology ; Animals ; Cattle/*embryology/physiology ; Cell Fusion ; Electroporation/veterinary ; Embryonic and Fetal Development/*drug effects ; Enzyme Inhibitors/administration & dosage/pharmacology ; Female ; Inositol 1,4,5-Trisphosphate/administration & dosage/*pharmacology ; Microinjections/veterinary ; Nuclear Transfer Techniques ; Oocytes/drug effects/growth & development ; Parthenogenesis/*drug effects ; Protein Kinase Inhibitors ; Ryanodine/administration & dosage/*pharmacology ; Skin/cytology

The authors tried to evaluate the effect of the growth factors on cell proliferation in the human retina astrocyte culture. Each growth factor, EGF, bFGF, GGF, IGF-1 or PDGF, together with BrdU was added to astrocytes-enriched cultures in the serum-free media. The proliferating effect of each growth factor was evaluated by a double immunofluo- rescenece labelling for GFAP and BrdU. By and addition of each growth factor, the number of retinal astrocytes at the synthetic stage significantly increased than those of control group (p<0.01). In comparision, PDGF was more potent than IGF-1 (p<0.01). Above data extended the concept of astrocytic proliferation caused by several growth factors in human retinal injury.
Astrocytes ; Bromodeoxyuridine ; Cell Proliferation ; Culture Media, Serum-Free ; Epidermal Growth Factor ; Humans ; Insulin-Like Growth Factor I ; Intercellular Signaling Peptides and Proteins* ; Neuroglia* ; Retina ; Retinaldehyde*